Exposure Experiments to Test the Kinetic Stability of 5‐Hydroxymethylfurfural Oxidase (HMFO) in Different Reactor Environments

ChemCatChem, vol. 17

Abstract

Abstract The impact of agitation on protein aggregation is often misattributed to shear stress rather than related phenomena such as cavitation and gas entrainment from the surface. For some time now, it has been known that shear is unlikely to harm most proteins directly. Rather, interfacial phenomena, particularly those involving dynamic gas‐liquid interfaces are critical contributors to protein damage, which leads to aggregation and compromises stability. This work investigated the kinetic stability of 5‐hydroxymethylfurfural oxidase (HMFO; EC: 1.1.3.47) in a 2 L stirred tank reactor. Exposure experiments revealed that the leading cause of enzyme deactivation was exposure to the gas‐liquid interface, either produced deliberately when sparging gas into the system or by accidental air entrainment from the overhead space due to mechanical stirring. This was further proven by experiments using the Bio Thrust membrane module, which enabled bubble‐free aeration thus, confirming that exposure to the gas‐liquid interface is the leading cause of deactivation.

Authors 5

1. Amalie Vang Høst

   AlbaNova · KTH Royal Institute of Technology · Technical University of Denmark

   Affiliation as printed

   Department of Chemical and Biochemical Engineering Technical University of Denmark (DTU) 2800 Kgs. Lyngby Denmark

   Department of Industrial Biotechnology School of Engineering Science in Chemistry Biotechnology, and Heath Royal Institute of Technology (KTH) Stockholm 11421 Sweden

   Royal Institute of Technology: Kungliga Tekniska Hogskolan Industrial Biotechnology AlbanNova Roslagstullsbacken 21 11421 Stockholm SWEDEN

2. Moritz Meyer Aachen

   RWTH Aachen University

   Affiliation as printed

   BioThrust Gmbh 52074 Aachen Germany

   Aachen University of Technology: Rheinisch-Westfalische Technische Hochschule Aachen BioThrust Gmbh Pauwelsstr. 17 52074 Aachen GERMANY

3. Patrick Bongartz Aachen

   RWTH Aachen University

   Affiliation as printed

   BioThrust Gmbh 52074 Aachen Germany

   Aachen University of Technology: Rheinisch-Westfalische Technische Hochschule Aachen BioThrust Gmbh Pauwelsstr. 17 52074 Aachen GERMANY

4. Marco W. Fraaije

   University of Groningen

   Affiliation as printed

   Molecular Enzymology University of Groningen Groningen 9747AG Netherlands

   University of Groningen: Rijksuniversiteit Groningen Molecular Enzymology Nijenborgh 3 97474G Groningen NETHERLANDS, KINGDOM OF THE

5. John M. Woodley corresponding

   Technical University of Denmark

   Affiliation as printed

   Department of Chemical and Biochemical Engineering Technical University of Denmark (DTU) 2800 Kgs. Lyngby Denmark

   Technical University of Denmark Department of Chemical Engineering S�ltofts Plads DK-2800 Lyngby DENMARK